JPH0772162B2 - Physiologically tolerated complex salt - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The objects of the invention are set forth in the appended claims.

Complexes or salts thereof have long been used in medicine, for example as an aid for the administration of sparingly soluble ions (eg iron) and for the detoxification of heavy metals or their radioisotopes by mistaken introduction (calcium in this case). Complex or zinc complex is superior).

By the way, according to the present invention, 13,23-dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontanedioic acid, trans-1,2-cyclohexylenediamine- N, N, N ', N'-tetraacetic acid, triethylenetetramine-N, N, N', N ", N, N-hexaacetic acid, 1,10-diaza-4,7-dioxadecane-1,1, 10,10−
Tetraacetic acid, 1,4,7,10-tetraazacyclododecane-N,
N ', N ", N-tetraacetic acid, tetraethylenepentamine-N, N, N', N", N, N ^IV , N ^IV -heptaacetic acid and diethylenetriamine-N, N, N ', N ", N A complexing acid anion selected from "-penta (methanephosphonic acid), an ion of a nonradioactive paramagnetic element having an atomic number of 21 to 29, 42, 44 or 57 to 83, and a first, second or second Physiologically tolerable complex salts of organic bases of 3 amines or at least one physiologically non-hazardous cation of lysine, arginine or ornithine are diagnostic agents suitable for NMR-, X-ray- or ultrasonic diagnostic methods. It proved to be very suitable for production.

The elements of the above atomic numbers forming the central ion of the physiologically tolerated complex salt must not be radioactive for the intended use of the diagnostic agent, of course.

If the agent containing the complex salt according to the invention is intended for use in NMR-diagnostics (European patent application no.
71564 (see JP-A-59-29718), the central ion of the complex salt must be paramagnetic. This is especially the atomic number
It is a divalent and trivalent ion of the elements 21-49, 42, 44 and 58-70. Suitable ions are, for example, chromium (III)-,
Manganese (II)-, Iron (III)-, Iron (II)-, Cobalt (II)-, Nickel (II)-, Copper (II)-, Praseodymium (III)-, Neudim (III)-, Samarium ( II
I)-and ytterbium (III) -ions. Owing to its extremely strong magnetic moments, gadolinium (III)-, terbium (III)-, dysprosium (II)
I)-, Holmium (III)-and Erbium (III)
-Ions are excellent.

If the complex-containing agent according to the invention is intended for use in X-ray diagnostics, the central ion must be derived from a higher atomic number element in order to achieve sufficient absorption of X-rays. I won't. Diagnostic agents containing physiologically tolerated complex salts with central ions of the elements of atomic numbers 57 to 83 have been found to be suitable for this purpose. These are, for example, lanthanum (III) -ions, the aforementioned ions in the lanthanide series, gold (III) -ions, lead (II) -ions or especially bismuth (III) -ions.

Both the agents for use in NMR-diagnostics and the agents for use in X-ray diagnostics are suitable for use in ultrasound-diagnostics.

The complexing acid is bound as a conjugate with biomolecules known to be abundant in the organ or organ site to be examined. Such biomolecules are eg hormones such as insulin, prostaglandins, steroid hormones, amino sugars, peptides, proteins or lipids. Conjugates with albumin, eg human serum albumin, antibodies, eg monoclonal antibodies specific for tumor-bound antigens or anti-myosin. Diagnostic agents formed therefrom are suitable for use in eg diagnostic methods for tumors and infarctions. For liver tests, conjugates with liposomes or clathrate compounds used as, for example, unilamellar or multilamellar phosphatidylcholine cholesterol vesicles are suitable. When the conjugate formation of or Protein or peptide through the carboxyl group of the complexing acid is, for example, based _{on: (CH 2) p -C 6} H 4 -W-
(P is 0 or 1, W is -NN-, -NHCS- or -NH
COCH ₂ - through a representative) are performed. In complex formation of a complexing acid with a protein, peptide or lipid, a large number of acid residues may partially bind to a macromolecule. In this case, the complexing acid residues each have a central ion. If the complexing acid is not bound to the biomolecule, the acid optionally has two, especially one central ion.

When all the acid hydrogen atoms of the complexing acid are not replaced by one or several central ions, the remaining hydrogen atoms are added to the organic base or amino acid physiologically to increase the solubility of the acid. It is advantageous to replace it by a non-hazardous cation. Suitable cations of organic bases are among others primary, secondary or tertiary amines such as ethanolamine, diethanolamine, morpholine, glucamine, N,
N-dimethylglucamine or especially N-methylglucamine.

The complexing acid required for the drug containing the complex salt according to the present invention is known or can be produced by a method known per se.

For example, the preparation of 13,23-dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontanedioic acid was carried out by the method proposed by RA Bulman et al. "NATO Abissen Shakhten (Na
turwissenschaften) ", Vol. 68, 483 (1981)]: 1,5-bis (2,6-dioxomorpholino) -3-azapentane-3-acetic acid 17.85 g (50 mmol) Suspended in 400 ml of anhydrous dimethylformamide and after addition of 20.13 g (100 mmol) of 11-aminoundecanoic acid are heated for 6 hours at 70 ° C. The clear solution is concentrated in vacuo, the yellow oily residue with 500 ml of water. Stir at room temperature, whereupon an almost white, voluminous solid precipitates, which is filtered off with suction and washed several times with water, the product obtained is taken up in 200 ml of acetone for further purification, After stirring at room temperature for 30 minutes, 36.9 g (97% of theory) of a white powder with a melting point of 134 ° -138 ° C. are obtained after suction filtration and drying at 50 ° C. in a vacuum.

The binding of the complexing acid to the biomolecule is likewise carried out in a manner known per se, for example by reacting the nucleophilic group of the biomolecule, such as an amino, hydroxy, thio or imidazole group, with an activated derivative of the complexing acid. Performed by.

As the activated derivative of the complexing acid, for example, acid chloride,
Mention may be made of anhydrous acids, activated esters, nitrenes or isothiocyanates. Conversely, it is likewise possible to react the activated biomolecule with a complexing acid.

Structure for binding to protein: −C ₆ H ₄ N ₂ ⁺ or C ₆ H ₄ NHC
Substituents for OCH ₂ halogen are also proposed.

The preparation of the complex salts is partially likewise known or in a manner known per se, in the manner of metal oxides or salts of the elements with atomic numbers 21 to 29, 42, 44 or 57 to 83 (for example nitrates, oxides or sulphates). Salt) in water and / or lower alcohol (eg methanol, ethanol or isopropanol) and add complexing acid equivalent solution or suspension in water and / or lower alcohol, and required By heating to the boiling point or stirring with heating until the reaction is completed. If the complex salt formed is insoluble in the solvent used, it is isolated by filtration. If soluble, the solution can be isolated by evaporation to dryness, for example by spray drying.

If the acid salt is present in the complex salt obtained, the acidic complex salt is converted into a neutral complex salt by using an inorganic and / or organic base or an amino acid which forms a physiologically harmless cation, and It is often advantageous to isolate this.
In many cases this is even unavoidable, since the dissociation of the complex salt is suppressed by the transfer of the pH value to neutrality, which only allows the isolation of the monolithic product, or at least the purification. Is.

The preparation is preferably carried out with organic bases or basic amino acids. However, it is also advantageous to carry out the neutralization with an inorganic base of sodium, potassium or lithium (hydroxide, carbonate or bicarbonate).

For the production of the neutral salt, the desired base may be added to the acidic complex salt of the aqueous solution or suspension in an amount reaching the neutralization point. The resulting solution can subsequently be concentrated to dryness in vacuo. Addition of a solvent capable of mixing the formed neutral salt with water, such as lower alcohol (methanol, ethanol, isopropanol, etc.), lower ketone (acetone, etc.), polar ether (tetrahydrofuran, dioxane, 1,2-dimethoxyethane, etc.) It is often advantageous to obtain a crystalline product which is precipitated by and thus is easy to isolate and of good purification.
It has proved to be particularly advantageous to add the desired base already during the complexation of the reaction mixture and thereby omit one step.

When the acidic complex salt contains several free acid groups, it is often advantageous to prepare a neutral mixed salt containing both inorganic and organic physiologically harmless cations as counterions. This involves reacting, for example, a complexing acid in an aqueous suspension or solution with an oxide or salt of the element providing the central ion and an amount of organic base which is 1/2 the amount necessary for neutralization, The complex salt formed is isolated, optionally purified and then added with the required amount of an inorganic base for complete neutralization. The order of base addition may be reversed.

The preparation of the diagnostic agents containing the complex salts according to the invention is likewise carried out in a manner known per se, by suspending or dissolving the complex salts in an aqueous medium, optionally with the addition of additives customary in Gasnus Pharmaceutical, and subsequently in solution or suspension. Sterilize the liquid. Suitable additives are, for example, physiologically non-hazardous buffers (eg tromethamine hydrochloride), small additions of complexing agents (eg diethylenetriamine-pentaacetic acid) or, if necessary, electrolytes (eg sodium chloride).

It is also possible in principle to produce the diagnostic agent without isolation of the complex salts according to the invention. In each case, special care must be taken to carry out the chelation in such a way that the salts and salt solutions according to the invention are not complexed and are substantially free of toxic metal ions. This can be ensured, for example, during the manufacturing process by control titration with a dye indicator, for example Zylenol Orange. Therefore, the present invention also relates to a method for producing a complex compound and a salt thereof. Purification of the isolated complex salt is the final guarantee.

If a suspension of the complex salt in water or physiological salt solution is desired for oral administration or other purposes, the slightly soluble complex salt may be replaced with one or more auxiliary agents commonly used in Galenian Pharma and / or surfactants. And / or mixed with aromas for taste correction.

The diagnostic agent is preferably 1 mmol to 1 mmol of the complex salt according to the invention.
It contains 1 mol and is usually dosed in an amount of 0.001 to 5 mmol / kg. It is suitable for oral administration and especially for parenteral administration.

The complex-containing agents according to the invention fulfill various prerequisites for their suitability as contrast agents in nuclear spin tomography. The drug is highly suitable for improving the testimony of the images obtained on nuclear spin tomography by increasing the signal intensity after oral or parenteral administration. In addition, the drug exhibits the high potency required to minimize the amount of xenobiotics that burden the body and the good compatibility required to maintain the non-invasive nature of the test ["J. Comput.Tomography ", 5 , 6 ,:
5 43-46 (1981, "Radiology", 144,343 (1982)
And "Brevet Special de Medicament", No.484M (19
The compounds listed in 1960 have strong toxicity, for example]. The good water solubility of the agents containing the complex salts according to the invention makes it possible to produce highly concentrated solutions, thus keeping the volume loading of the circulation within an acceptable range and adjusting the dilution with body fluids. That is, the NMR-diagnostic agent must be 100 to 1000 times more water soluble than that associated with NMR spectroscopy. Furthermore, the drug not only exhibits a high stability in vitro but also a surprisingly high stability in vivo, so that the release or exchange of the non-covalently bound self-toxic ion within the complex is It occurs very slowly within 24 hours when the contrast agent is completely redeposited (as pharmacological studies have shown). Conjugates with proteins and antibodies, which are used, for example, in tumor diagnostics, already give surprisingly high signal amplifications in very low amounts, so that in this case also correspondingly low concentrations of solution can be used. .

It should be particularly emphasized that the agents containing the complex salts according to the invention are very suitable as X-ray contrast agents, in which the signs of anaphylactic reaction known for iodine-containing contrast agents are not observed in biochemical-pharmacological examinations. is there. The drug is particularly useful for its advantageous absorption in the higher tube voltage range for digital subtraction techniques.

The agent containing the complex salt according to the present invention is also suitable as an ultrasonic diagnostic agent on the basis of its property of acting favorably on the ultrasonic velocity.

Unlike conventional X-ray diagnostics which use X-ray contrast agents to give a shadow picture, there is no linear dependence of signal amplification on the concentration used in NMR-diagnostics which use paramagnetic contrast agents. Higher doses do not unconditionally lead to signal amplification as control studies show, and higher doses of paramagnetic contrast agents can lead to signal loss. For this reason, some pathological processes are described in European Patent Application No. 71564 (= Japanese Patent Application No. 57-1278).
No. 10) The dose stated in the specification (this is 0.001 mmol /
It was surprising that visualization was possible only after the administration of a contrast agent containing a ferromagnetic complex according to the invention of a dose higher than (kg to 5 mmol / kg). For example, the diagnosis of blood-brain barrier in head tumor failure is paramagnetic complex salts, such as gadolinium-diethylenetriaminepentaacetic acid or manganese-1,2-cyclohexylenediamino-tetraacetic acid 0.05-2.5 mmol / kg, in the form of readily water-soluble salts. Advantageously 0.
Only obtained after administration of 1-0.5 mmol / kg. Higher concentrations of up to 1 mol / l, preferably 0.25 to 0.75 mol / l, are required for doses above 0.1 mmol / kg, since this alone reduces the volume loading and the injection solution. Is guaranteed.

Particularly low doses (below 1 mg / kg) and thus lower concentrated solutions (1 μmol / l to 5 mmol // l) than those described in the above-mentioned JP-A-57-127810 are for example tumors and myocardium. Requirement in organ-specific NMR-diagnostics for infarct detection.

Next, the present invention will be described in detail with reference to Examples.

Example 1 (Reference Example) nitrilo -N, N, of N- triacetate gadolinium (III) - preparation of complexes (C ₆ H ₆ GdNO ₆₎ of gadolinium oxide in water _{1.2l (Gd 2 O 3) 36.2g} (100 Mmol) and 38.2 g (200 mmol) of nitrilotriacetic acid are heated with stirring to 90 ° -100 ° C. and stirred for 48 hours at this temperature. Impurities are then filtered off via activated carbon and the filtrate is evaporated to dryness. Powder the amorphous residue.

Yield: 60 g; 87% of theory. Melting point: 300 ° C. Gadolinium: Calculated 45.5%, Found 44.9%.

Example 2 A) Reference Example 13,23-Dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontane-Diacid of a gadolinium (III) complex of diacid. sodium salt preparation _{(C 36 H 60 GdN 5 O} 12 · 2Na) 13,23- dioxo -15,18,21- tris (carboxymethyl) -12,15,18,21,24- pentaaza pentatriacontanoic Con Tan - 15.2 g (20 mmol) of the diacid are suspended in 400 ml of water and heated to 95 ° C. Gadolinium (II dissolved in 60 ml of water
I) -Chloride hexahydrate (7.43 g, 20 mmol) is gradually added dropwise. Hold at this temperature for 2 hours and subsequently add 60 ml of 1N caustic soda solution to neutralize the hydrochloric acid formed.

Complete reaction (tested with Zylenol Orange) The precipitate obtained is filtered and washed with chloride until free of chloride. 17.60 g of water-insoluble white powder with a melting point of 290-292 ℃ (theoretical
96%) is obtained.

Gadolinium (III) complex of 13,23-dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontane-diacid Analysis: Calculated: C47.30 H6.84 N7.66 Gd17.20 Found: C47.13 H6.83 N7.60 Gd17.06 14.6 g (16 gadolinium (III) -complex thus obtained
Millimole) in 200 ml of water and 31.4 ml of 1N caustic soda solution are added in small portions. After 1 hour a clear solution is obtained, which is filtered and subsequently concentrated in vacuo. In vacuum
After drying at 80 ° C, easily water-soluble with a melting point of 279-285 ° C-white powder 13.2
g (87% of theory) is obtained.

Analysis: Calculated value: C45.13 H6.31 N7.31 Gd16.41 Na4.80 Measured value: C45.20 H6.12 N7.28 Gd16.26 Na4.75 Relaxability (20MHz, 39 ° C) T ₁ : 5.72 T ₂ : 6.84 (L / mmol · sec) LD ₅₀ (intravenous, rat)> 1.52 mmol / kg B) In the same manner as the present invention, N-methylglucamine was used instead of caustic soda solution to obtain 13,23-dioxo- Di- of gadolinium (III) complex of 15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontane-diacid
The N-methylglucamine salt (C ₅₀ H ₉₆ GdN ₇ O ₂₂ ) is obtained as a white powder with a decomposition point above 30 ° C.

Analysis (for anhydrous substance): Calculated value: C46.03 H7.42 N7.52 Gd12.05 Found value: C46.22 H7.61 N7.44 Gd11.92 Example 3 A) Reference example Diethylenetriamine-N, N, N ', N ", N" - preparation of di -N- methylglucamine salt of gadolinium (III) complex of pentaacetic acid _{(C 28 H 54 GdN 5 O} 20) diethylenetriamine -N, N, N', N ", N ″ -Pentaacetic acid 3
9.3 g (100 mmol) are suspended in 200 ml of water and 19.5 g (100 mmol) N-methylglucamine are added. 18.12 g (50 mmol) of gadolinium (III) oxide (Gd ₂ O ₃ ) are subsequently added in small portions and the resulting suspension is heated to 95 ° C. About 1 hour later N-methylglucamine 19.5 g (1
(00 mmol) is added and after heating for a further 2 hours a clear solution is obtained. After complete reaction (control with Zylenol Orange) a small amount of insoluble material is filtered off and the filtrate is concentrated to dryness in vacuo. The residue is freshly dissolved in 100 ml of water and stirred into 250 ml of ethanol. After cooling for several hours, the crystalline product is suction filtered, washed with cold ethanol and dried at 60 ° C. in vacuo. 92.7 g (99% of theory) of a white powder with an unspecified melting point are obtained.

Analysis: Calculated value: C35.85 H5.80 N7.47 Gd16.77 Actual value: C35.50 H5.72 N7.20 Gd16.54 Relaxability (20MHz, 39 ° C) T ₁ : 3.71 T ₂ : 4.09 (L / mmol · sec) LD ₅₀ (intravenously, rat) about 9 mmol / kg B) in the same manner as the present invention example a, triethylenetetramine -N with gadolinium oxide, the _{Gd 2 O 3, N, N} ' , N ″, N, N−
The tri-N-methylglucamine salt of the gadolinium (III) complex of hexaacetic acid, C ₃₉ H ₇₈ GdN ₇ O ₂₇ is obtained: Analysis (for anhydrous substance): Calculated: C37.95 H6.37 N7.94 Gd12 .74 Found: C37.81 H6.50 N7.88 Gd12.53 LD ₅₀ (intravenous, mouse): ca. 5 mmol / kg This salt occurs as a white powder with an unspecified melting point. They are very well soluble in water.

Example 4 A) Reference Example Preparation of disodium salt of gadolinium (III) complex of diethylenetriamine-N, N, N ', N ", N" -pentaacetic acid (C
₁₄ H ₁₈ GdN ₃ O ₁₀・ 2Na) gadolinium (III) oxide 18.2 g (0.05 mol) and diethylenetriamine-pentaacetic acid 39.3 g (0.1 mol) were added to water 11
Suspend in 0 ml and heat to reflux for 1 hour. Cool the clear solution and add about 80 ml of 5N-caustic soda solution to pH 7.5.
To Heat to a new boil and add 250 ml of ethanol dropwise. After stirring for a few hours in an ice bath, the crystalline product is suction filtered, washed with ice-cold ethanol and dried at 60 ° C. in vacuo. A white powder is obtained in quantitative yield, which does not melt up to 300 ° C.

Analysis: Calculated value: C28.43 H3.07 N7.10 Gd26.58 Measured value: C28.35 H2.95 N7.05 Gd26.37 B) Reference example Using gadolinium (III) oxide and Gd ₂ O ₃ tetra Ethylene pentamine-N, N, N ', N ", N, N ^IV , N ^IV- Sodium salt of digadolinium (III) complex of heptaacetic acid, C ₂₂ H ₃₀ Gd ₂ N ₅ O ₁₄ · Na; Analysis ( For anhydrous substances): Calculated: C28.54 H3.27 N7.56 Gd33.96 Found: C28.32 H3.40 N7.66 Gd33.78 This salt occurs as a white powder with an unspecified melting point,
Very well water-soluble.

Example 5 A) Reference example N of iron (III) complex of diethylenetriaminepentaacetic acid
-Production of methylglucamine salt (C ₂₁ H ₃₇ FeN ₄ O ₁₅ ) 35.40 g (90 mmol) diethylenetriaminepentaacetic acid
Was suspended in 100 ml of water and dissolved in 100 ml of water. 24.3 g (90 mmol) of iron (III) chloride hexahydrate (FeCl ₃ .6H ₂ O)
Add. The initially dark brown suspension is heated to 95 ° C. After about 1 hour, this color changes to light yellow. 270 ml of 1N caustic soda solution was added to neutralize the generated hydrochloric acid, and further 3
Heat to 95 ° C for hours. The bright yellow precipitate obtained is suction filtered, washed free of chloride with water and dried at 60 ° C. in vacuo. 17.85 g (45% of theory) of a light yellow powder with a melting point> 300 ° C. are obtained.

17.85 g (40 mmol) of the obtained iron (III) complex was added to 200 ml of water.
Solid N-methylglucamine little by little.
7.8 g (40 mmol) are added. Heat to 50 ° C for about 3 hours
And an almost clear reddish brown solution is obtained, which is filtered and then concentrated to dryness in vacuo. The residue is dried in vacuum at 50 ° C. 24.3 g (95% of theory) of reddish brown powder with a melting point of 131-133 ° C.
%) Is obtained.

Analysis: Calculated value: C39.82 H5.89 N8.85 Fe8.81 Measured value: C39.70 H6.00 N8.65 Fe9.01 B) The present invention correspondingly using N-methylglucamine Is obtained: trans-1,2-cyclohexylenediamine-N, N, N ′,
Of N'- tetraacetic acid iron (III) complex N- methylglucamine _{salt, C 21 H 36 FeN 3 O} 13; Analysis (relative to anhydrous substance): Calculated: C42.44 H6.10 N7.07 Fe9.40 Found: C42.40 H6.30 N7.20 Fe9.21 Tri-N-methylglucamine salt of iron (III) complex of triethylenetetramine-N, N, N ', N ", N, N-hexaacetic acid. , C ₃₉ H ₇₈ FeN ₇ O ₂₇ ; Analysis (for anhydrous substances): Calculated value: C41.36 H6.94 N8.66 Fe4.90 Measured value: C41.22 H6.99 N8.72 Fe4.81 Example 6 ( The present invention) trans-1,2-cyclohexylenediamine-N, N, N ',
N'- manufacture of gadolinium (III) complex of N- methylglucamine salt of tetraacetic acid _{(C 21 H 36 GdN 3 O} 13) trans-1,2-cyclohexylene-diamine -N, N, N ',
20.78 g (60 mmol) of N'-tetraacetic acid are suspended in 150 ml of water. After the addition of 11.7 g (60 mmol) N-methylglucamine, an almost clear solution is obtained, to which 10.88 g (30 mmol) gadolinium oxide (Gd ₂ O ₃ ) is added. The freshly obtained suspension is heated to 75 ° C. for 6 hours. A small amount of insoluble matter is filtered off, and the filtrate is concentrated to dryness. Residue in vacuum at 60 ° C
Dry and powder. White powder with a melting point of 258-261 ℃
38.6 g (92% of theory) are obtained.

Analysis: Calculated value: C36.25 H5.22 N6.04 Gd22.60 Actual value: C36.40 H5.50 N5.98 Gd22.52 Relaxability (20MHz, 39 ° C) T ₁ : 6.13 T ₂ : 7.13 (L / Mmol · sec) LD ₅₀ (intravenous, rat) about 2 mmol / kg Example 7 A) Reference example trans-1,2-cyclohexylenediamine-N, N, N ′,
N'- preparation of disodium salt of tetraacetic acid manganese (II) complex _{(C 14 H 18 MnN 2 O} 8 · 2Na) trans-1,2-cyclohexylene-diamine -N, N, N ',
34.6 g (100 mmol) of N'-tetraacetic acid was added to 100 parts of water under nitrogen.
and manganese (II) carbonate, MnCO ₃ 11.5g
(100 mmol) is added. Heat to 95 ° C and add 200 ml of 1N caustic soda solution in small portions. The clear solution is concentrated in vacuo and the residue is dried in vacuum at 60 ° C. 40.8 g (92% of theory) of a rose-colored powder are obtained.

Analysis: Calculated value: C37.94 H4.09 N6.32 Mn12.40 Measured value: C37.78 H4.12 N6.20 Mn12.31 Relaxability (20MHz, 39 ° C) T ₁ : 2.94 T ₂ : 5.03 (L / mmol · sec) LD ₅₀ (intravenous, rat): the following to be obtained with N- methylglucamine instead of about 8 mmol / kg B) present invention sodium hydroxide solution: trans-1,2-cyclohexylene cyclohexylene diamine - di -N- methylglucamine salt of the manganese (II) complexes of tetra _{acetate, C 28 H 54 MnN 4 O} 18; analysis (relative to anhydrous substance): calculated: C42.59 H6.89 N7.09 Mn6 .96 Found: C42.40 H6.95 N7.20 Mn7.18 Relaxability (20MHz, 39 ° C) T ₁ : 2.63 T ₂ : 4.00 (L / mmol · sec) LD ₅₀ (intravenous, rat): Approx. 15 mmol / kg Example 8 A) Reference example N-methylglucamine salt of gadolinium (III) complex of ethylenediamine-N, N, N ', N'-tetraacetic acid (C ₁₇ H ₃₀ G
dN ₃ O ₁₃₎ ethylenediamine -N, N, N ', N'- tetraacetic acid 29.2 g (10
0 mmol) is suspended in 100 ml of water and heated to 95 ° C. with 18.1 g (50 mmol) of gadolinium (III) oxide. During the heating, 19.5 g (100 mmol) of N-methylglucamine are added in small portions. After about 3 hours a clear solution is obtained, which is filtered and concentrated to dryness in vacuo. The rest
Dry in vacuum at 60 ° C. 61.3 g (95% of theory) of a white powder with an unspecified melting point are obtained.

Analysis: Calculated value: C31.82 H4.71 N6.55 Gd24.51 Actual value: C31.65 H4.59 N6.52 Gd24.56 B) The following can be obtained in the same manner as the present invention: 1,10- N-methylglucamine salt of gadolinium (III) complex of diaza-4,7-dioxadecane-1,1,10,10-tetraacetic acid, C ₂₁ H ₃₈ GdN ₃ O ₁₅ ; Analysis (with respect to anhydrous substance): Calculated value : C34.56 H5.25 N5.76 Gd21.55 Actual value: C34.38 H5.40 N5.55 Gd21.30 Example 9 A) Reference example 1,4,7,10-tetraazacyclododecane-N, N ′, N ″, N
-Preparation of sodium salt of gadolinium (III) complex of tetraacetic acid (C ₁₆ H ₂₄ GdN ₄ O ₈ · Na) 1,4,7,10-Tetraazacyclododecane-N, N ′, N ″, N
-Suspending 4.0 g (10 mmol) of tetraacetic acid in 20 ml of water,
And add 10 ml of 1N-caustic soda solution. Gadolinium (III) oxide, 1.8 g (5 mmol) Gd ₂ O ₃ are added and the suspension is heated to 50 ° C. for 2 hours. The clear solution is filtered and concentrated in vacuo. The residue is dried and powdered. 5.5 g (95% of theory) of a white powder are obtained.

Analysis: Calculated value: C33.10 H4.17 N9.65 Gd27.08 Measured value: C33.01 H4.20 N9.57 Gd27.16 Relaxability (20MHz, 39 ° C) T ₁ : 1.73 T ₂ : 2.16 (L / mmol · sec) LD ₅₀ (intravenous, mouse): about 7.5 mmol / kg B) the invention Similarly the following to be obtained: 1,4,7,10-tetraazacyclododecane -N, N ' , N ″, N
- tetra acetic acid gadolinium (III) complex N- methylglucamine _{salt, C 23 H 42 GdN 5 O} 13; Analysis (relative to anhydrous substance): Calculated: C36.64 H5.62 N9.29 Gd20.86 Found : C36.58 H5.70 N9.11 Gd20.95 Example 10 A) Reference example Ethylenedinitrilo-tetrakis (methanephosphonic acid)
Gadolinium (III) preparation of tetra -N- methylglucamine salt of the complex _{(C 34 H 85 GdN 6 O} 32 P 4) ethylene dinitrilo - tetrakis (methanephosphonic acid)
9.11 g (20 mmol) are suspended in 150 ml of water and brought to pH 5 with the corresponding amount of N-methylglucamine. Add gadolinium (III) oxide, 3.6 g (10 mmol) Gd ₂ O ₃ and heat to 70 ° C. After about 1 hour, a clear solution is obtained, to which the remaining amount of N-methylglucamine is added. A total of 15.6 g (80 mmol) of N-methylglucamine was consumed. The solution is concentrated to dryness in vacuo and the remaining gel residue is charged to 200 ml of acetonitrile. About 20 hours at 30 ℃
And the fine residue obtained is filtered off with suction. 40
23.4g of white powder with melting point of 115-118 ℃ after drying in vacuum at ℃
(85% of theory) is obtained.

Analysis: Calculated value: C29.78 H6.25 N6.13 P9.04 Gd11.47 Actual value: C29.85 H6.57 N5.98 P8.78 Gd11.26 B) Diethylenetriamine-N, N , N ′, N ″, N ″ −
Hepta-N-methylglucamine salt of gadolinium (III) complex of penta (methanephosphonic acid), C ₅₈ H ₁₄₄ GdN ₁₀ O
₅₀ P ₅ obtained; analysis (for anhydrous substance): Calculated: C33.27 H6.93 N6.69 Gd7.51 Found: C33.40 H7.05 N6.51 Gd7.40 Example 11 (reference example) Diethylenetriamine-pentaacetic acid gadolinium (II
I) Preparation of a mixture salt solution of sodium salt of complex and N-methylglucamine salt a) Preparation of mono-N-methylglucamine salt of complex, C
₂₁ H ₃₇ GdN ₄ O ₁₅ 195.2 g (1 mol) of N-methylglucamine salt is dissolved in 7 l of water. Next, 393.3 g of diethylenetriaminepentaacetic acid (1
Mol) and gadolinium oxide, 181.3 g of Gd ₂ O ₃ (0.5 mol) are added, and the mixture is heated under reflux for 2 hours. The filtered clear solution is spray dried. A white crystalline powder with a water content of 2.6% is obtained, which melts at 133 ° C. and melts at boiling at 190 ° C.

Gd: Calculated value 21.17 Measured value 21.34 b) Preparation of neutral mixed salt solution 730.8 g (1 mol) of the salt obtained in a) was added to water (pi) (Wass).
er pro injectione = distilled water for injection) 630 ml,
And 40 g (1 mol) of caustic soda powder is added little by little. Water (pi) is added to this neutral solution to make 1000 ml, which is filled in a bottle through a pyrogen filter and sterilized by heating. This 1M solution contains 753.8 g of mixed salt per day.

Example 12 (Invention) Preparation of a solution of the N-methylglucamine salt of a gadolinium (III) complex of 1,4,7,10-tetraazacyclododecanetetraacetic acid 370.9 g (500 mmol) of the salt mentioned in Example 9B Water (p.
i.) Suspend in 500 ml and add water (pi) to dissolve to make 1000 ml. The solution is placed in an ampoule and sterilized by heat.

Example 13 (Invention) Preparation of a solution of the di-N-methylglucamine salt of the manganese (II) complex of trans-1,2-cyclohexylenediaminetetraacetic acid 395.9 g (500 mmol) of the salt mentioned in Example 7B (Pi) 500
Resuspend in ml. Add 1.3 g of ascorbic acid, and add water (pi) to make 1000 ml, and make a solution. The solution is sterile filtered and filled into ampoules.

Example 14 (Reference example) Preparation of solution of di-N-methylglucamine salt of iron (III) complex of diethylenetriamine-pentaacetic acid 44.6 g (0.1%) of iron (III) complex of diethylenetriamine-pentaacetic acid obtained in Example 5A Mol) in 40 ml of water (pi). After adding 0.18 g of tromethamine-hydrochloride and 39.1 g (0.2 mol) of N-methylglucamine, the mixture was neutralized and dissolved, and water (pi) was added to this solution to make 100 ml,
Fill ampoules and heat sterilize.

Example 15 (Reference example) Preparation of solution of gadolinium (III) complex of nitrilotriacetic acid 1.9 g (10 mmol) of nitrilotriacetic acid and 1.8 g (5 mmol) of gadolinium (III) oxide are dissolved in 100 ml of water (pi) under heating. The solution is made into an ampoule and sterilized by heat.

Example 16 (Invention) Preparation of a solution of N-methylglucamine salt of gadolinium (III) complex of trans-1,2-cyclohexylenediamine-tetraacetic acid 555.8 g (0.8 mol) of the salt described in Example 6 was added to water ( pi) to 1000 ml. After filtration through a pyrogen filter, the solution is filled into ampoules and sterilized by heating.

Example 17 (Reference Example) N '-(2-hydroxyethyl) ethylenediamine-N,
Preparation of solution of gadolinium (III) complex of N, N'-triacetic acid N '-(2-hydroxyethyl) ethylenediamine-N,
1.9 g (6.7 mmol) N, N'-triacetic acid and 1.2 g (3.35 mmol) gadolinium oxide are dissolved in 6 ml water (pi) with heating. The solution is placed in an ampoule and sterilized by heat.

Example 18 (invention) 13,23-dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontanedioic acid gadolinium (III) complex di- Preparation of a solution of N-methylglucamine salt 130.4 g (100 mmol) of the salt listed in Example 2B was added to water (pi) 250
Suspend in ml and dissolve under heating. Add water (pi) to make 500 ml, put the solution in an ampoule, and sterilize by heat.

Example 19 (Reference Example) Production of Liposomes Containing Gadolinium-DTPA "Procedures of the National Academy of Sciences of the United States of America (Proc. .U.
SA) "[Vol. 75, p. 4194] by the treatment method described in [Egg-phosphatidylcholine 75 mol% and cholesterol.
A 25 mol% lipid mixture is prepared as dry substance.
From this 500 mg are dissolved in 30 ml diethyl ether and 3 ml of a 0.1 M solution of the di-N-methylglucamine salt of the gadolinium (III) complex of diethylenetriamine-pentaacetic acid in water (pi) is added in an ultrasonic bath. After complete addition of the solution, sonication is continued for another 10 minutes and then concentrated in a rotary evaporator. The gel residue is suspended in 0.125 mol-sodium chloride solution, and the contrast agent that is not encapsulated in liposomes is removed by centrifugation (20,000 kg / 20 minutes) at 0 ° C. The liposomes thus obtained are subsequently freeze-dried in a multi-glass bottle. Administration is carried out as a colloidal dispersion in 0.9% by weight saline solution.

Example 20 (Invention) 1,4,7,10-Tetraazacyclododecane-N, N ', N ", N
- production of lysine salt of the gadolinium (III) complex of tetra acetic acid _{(C 22 H 39 GdN 6 O} 10) 1,4,7,10- tetraazacyclododecane--N, N ', N ", N
40.0 g (100 mmol) of tetraacetic acid are suspended in 750 ml of water and mixed with 14.6 g (100 mmol) of lysine. 18.1 g (50 mmol) gadolinium (III) oxide Gd ₂ O ₃ are added and the suspension is heated to 50 ° C. for 5 hours. The clear solution is filtered and concentrated in vacuo. White powder 66.9g (9 in theory
5%) is obtained.

Analysis: Calculated: C37.49 H5.58 N11.92 Gd22.31 Found: C37.22 H5.66 N11.99 Gd22.14 Example 21 (invention) 1,4,7,10-tetraazacyclododecane −N, N ′, N ″, N
-Preparation of a solution of gadolinium (III) lysine salt of tetraacetic acid (DOTA) 211.4 g (300 mmol) of the salt listed in Example 20 was added to water (pi) 600
Make a paste in ml, mix with 500 mg DOTA and 500 mg sodium hydrogen carbonate and dissolve by adding water (pi) to 1000 ml. The solution is filled into ampoules and sterilized by heat.

Example 22 (Invention) 1,4,7,10-Tetraazacyclododecane-N, N ', N ", N
- preparation of di -N- methylglucamine salt of the manganese (II) complex of tetra acetic acid _{(C 30 H 54 MnN 6 O} 18) 1,4,7,10- tetraazacyclododecane--N, N ', N " , N
40.0 g (100 mmol) of tetraacetic acid are suspended in 400 ml of water and mixed with 39.0 g (200 mmol) of N-methylglucamine. Manganese carbonate MnCO ₃ 11.5 g (100 mmol)
Is added and the suspension is heated under nitrogen to 50 ° C. for 5 hours. The clear solution is filtered and concentrated in vacuo to give 75.8 g (90% of theory) of a pink powder.

Analysis: Calculated: C42.81 H6.47 N9.98 Mn6.53 Found: C42.66 H6.55 N9.88 Mn6.60 Example 23 (invention) 1,4,7,10-tetraazacyclododecane −N, N ′, N ″, N
-Preparation of a solution of iron (III) complex of tetraacetic acid 1,4,7,10-tetraazacyclododecane-N, N ', N ", N
20.0 g (50 mmol) of tetraacetic acid (DOTA) in water (p.
i.) Suspended in 1200 ml and sodium citrate C ₆ H ₅ Na
150 g of ₃ O _{7 and} 39.9 g of ferric oxide Fe ₂ O ₃ (25 mmol)
Mix with. The suspension is heated to 70 ° C. under nitrogen for 24 hours.
The clear solution is mixed with 300 g D-mannitol and 5 g DOTA and water (pi) is added to 2000 ml to dissolve and produce for oral application.

Example 24 (reference example) N, N ', N ", N-tetrakis (phosphonatemethyl)-
Pentameglumine salt of gadolinium (III) complex of 1,4,7,10-tetraazacyclododecane N, N ', N ", N-tetrakis (phosphonatemethyl) -1,4,7,10- in 500 ml of water Tetraazacyclododecane [“J.
Org.Chem. " 31 , 1603 (1966)] 274.15g (0.5
Gadolinium oxide Gd ₂ O ₃ 90.63 g (0.25 mol)
Mol) is added. Then, under heating, 488 g (2.5 mol) of N-methylglucamine (meglumine) was added little by little, and the mixture was stirred at room temperature for 24 hours. Acetone is added dropwise to the clear solution under cooperative stirring until turbidity begins and stirred for 48 hours in an ice bath. Precipitate, decant the mother liquor and again stir for 24 hours with 2.5 l of acetone at room temperature. The solid substance is filtered off with suction, washed with acetone and dried at 60 ° C. in a vacuum. The complex salt (753.1 g: 90% of theory) is obtained as a white powder with unspecified decomposition points.

Analysis (for anhydrous substance): Calculated: C33.73 H6.57 N7.53 P7.40 Gd9.40 Found: C33.85 H7.02 N7.48 P7.50 Gd9.51

[Brief description of drawings]

FIG. 1a corresponds to Example 9B of the present invention and was taken with the N-methylglucamine salt of the gadolinium (III) complex of 1,4,7,10-tetraazacyclododecane-tetraacetic acid (without Gd-DOTA). NMR contrast photograph of rat kidney, Fig. 1b is NMR contrast photograph of rat kidney taken using Gd-DOTA, and Fig. 2a is Gd-D.
An NMR contrast photograph of rat nephroma taken without OTA,
FIG. 2b is an NMR contrast photograph of rat nephroma taken using Gd-DOTA, and FIG. 3a is trans-1, which corresponds to Example 7B of the present invention.
Di-N-methylglucamine salt (Mn-CDTA) of manganese (II) complex of 2-cyclohexylenediamine-tetraacetic acid
NMR contrast photograph of rat kidney taken without Mn-CDTA, Fig. 3b is NMR contrast photograph of rat kidney taken with Mn-CDTA, and Fig. 4a is rat kidney taken without Mn-CDTA. NMR contrast photograph of tumor, FIG. 4b is an NMR contrast photograph of rat renal tumor photographed using Mn-CDTA.

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI Technical indication location C07F 13/00 A 9155-4H 15/00 Z 9155-4H (72) Inventor Hans-Joachim. Vwainmann Berlin, Germany 38. West Hohener. Wake 27 (56) Bibliography Chem. Abstr. 95 (8), 68865c (1981) Chem. Abstr. 91 (7), 52006e (1979)

Claims (5)

[Claims] 1. 13,23-Dioxo-15,18,21-tris (carboxymethyl) -12,15,18,21,24-pentaazapentatriacontanedioic acid, trans-1,2-cyclohexylenediamine -N, N, N ', N'-tetraacetic acid, triethylenetetramine-N, N, N', N ", N, N-hexaacetic acid, 1,10
-Diaza-4,7-dioxadecane-1,1,10,10-tetraacetic acid, 1,4,7,10-tetraazacyclododecane-N, N ', N ",
N-tetraacetic acid, tetraethylenepentamine-N, N,
A complexing acid anion selected from N ′, N ″, N ″, N ^IV , N ^IV -heptaacetic acid and diethylenetriamine-N, N, N ′, N ″, N ″ -penta (methanephosphonic acid), Atomic number 21-
From 29, 42, 44 or 57-83 non-radioactive paramagnetic element ions and at least one physiologically non-hazardous cation of an organic base of a primary, secondary or tertiary amine or lysine, arginine or ornithine Consisting of a physiologically tolerable complex salt. 2. The organic base of the primary, secondary or tertiary amine is ethanolamine, diethanolamine, morpholine, glucamine, N, N-dimethylglucamine or N-methylglucamine. Complex salt. 3. The complex salt according to claim 1, wherein the complexing acid is 1,4,7,10-tetraazacyclododecane-N, N ', N ", N-tetraacetic acid. 4. The complex salt according to claim 1, which is a di-N-methylglucamine salt of a manganese (II) -complex of trans-1,2-cyclohexylenediamine-tetraacetic acid. 5. The complex salt according to claim 1, which is an N-methylglucamine salt of a gadolinium (III) -complex of 1,4,7,10-tetraazacyclododecane-tetraacetic acid.